Counting and Breaking Single Bonds

Dynamic Force Spectroscopy in Tethered Single Molecule Systems
  • Todd A. Sulchek
  • Raymond W. Friddle
  • Aleksandr Noy

Tethered molecular systems, in which flexible polymer linkers connect the interacting molecules to the surfaces of the atomic force microscope probe and target sample, provide a particularly attractive platform for studying biological interactions using force spectroscopy. We describe the underlying nanomechanics of individual tether molecules. We provide a sampling from the literature that illustrates how knowledge and control of tether linkage can aid understanding of molecular interactions. We then describe the basic physical principles of force spectroscopy measurements of tethered biological interactions and show that well-defined mechanical properties of the tether linkages allow independent determination of the number of ruptured bonds. This approach allows us to show that forces between multiple biological bonds measured in a parallel configuration obey the predictions of a Markovian model of bond dissociation. Finally, we discuss the use of the dynamic force spectra of single and multiple protein-ligand bonds for determination of kinetic parameters for multivalent interactions. This ability to form, count, and dissociate biological bonds with nanomechanical forces provides a powerful method to study the physical laws governing the interactions of biological molecules.


Contour Length Bond Rupture Force Spectroscopy Rupture Force Tethered System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Cochran, J. R.; Cameron, T. O.; Stone, J. D.; Lubetsky, J. B.; Stern, L. J. J. Biol. Chem. 2001, 276, 28068–28074.CrossRefGoogle Scholar
  2. 2.
    van Kooyk, Y.; Figdor, C. G. Curr. Opin. Cell Biol. 2000, 12, 542–547.CrossRefGoogle Scholar
  3. 3.
    Mammen, M.; Choi, S. K.; Whitesides, G. M. Angew. Chem. 1998, International Ed. in English. 37, 2755–2794.Google Scholar
  4. 4.
    Mourez, M.; Kane, R. S.; Mogridge, J.; Metallo, S.; Deschatelets, P.; Sellman, B. R.; Whitesides, G. M.; Collier, R. J. Nat Biotech 2001, 19, 958–961.CrossRefGoogle Scholar
  5. 5.
    Holliger, P.; Prospero, T.; Winter, G. PNAS 1993, 90, 6444–6448.CrossRefADSGoogle Scholar
  6. 6.
    Todorovska, A.; Roovers, R. C.; Dolezal, O.; Kortt, A. A.; Hoogenboom, H. R.; Hudson, P. J. J. Immunol. Meth. 2001, 248, 47–66.CrossRefGoogle Scholar
  7. 7.
    Souriau, C.; Hudson, P. J. Expert Opin Biol Ther 2003, 3, 305–18.CrossRefGoogle Scholar
  8. 8.
    Rowland, G.; O’Neill, G.; Davies, D. Nature 1975, 255, 487–488.CrossRefADSGoogle Scholar
  9. 9.
    Bustamante, C.; Macosko, J. C.; Wuite, G. J. L. Nature Rev. Molec. Cell Biol. 2000, 1, 130–136.CrossRefGoogle Scholar
  10. 10.
    Clausen-Schaumann, H.; Seitz, M.; Krautbauer, R.; Gaub, H. E. Curr. Opin. Chem. Biol. 2000, 4, 524–530.CrossRefGoogle Scholar
  11. 11.
    Noy, A.; Vezenov, D.; Lieber, C. Ann. Rev. Mat. Sci. 1997, 27, 381–421.CrossRefADSGoogle Scholar
  12. 12.
    Merkel, R.; Nassoy, P.; Leung, A.; Ritchie, K.; Evans, E. Nature 1999, 397, 50–52.CrossRefADSGoogle Scholar
  13. 13.
    Alon, R.; Hammer, D. A.; Springer, T. A. 1995, 374, 539–542.Google Scholar
  14. 14.
    Thoumine, O.; Kocian, P.; Kottelat, A.; Meister, J.-J. Eur. Biophys. J.l 2000, 29, 398–408.CrossRefGoogle Scholar
  15. 15.
    Florin, E. L.; Moy, V. T.; Gaub, H. E. Science 1994, 264, 415–417.CrossRefADSGoogle Scholar
  16. 16.
    Moy, V. T.; Florin, E. L.; Gaub, H. E. Science 1994, 266, 257–9.CrossRefADSGoogle Scholar
  17. 17.
    Evans, E.; Ritchie, K. Biophys. J. 1997, 72, 1541–1555.CrossRefGoogle Scholar
  18. 18.
    Dietz, H.; Rief, M. Proc. Natl. Acad. Sci. U.S.A. 2004, 101, 16192–16197.CrossRefADSGoogle Scholar
  19. 19.
    Riener, C. K.; Stroh, C. M.; Ebner, A.; Klampfl, C.; Gall, A. A.; Romanin, C.; Lyubchenko, Y. L.; Hinterdorfer, P.; Gruber, H. J. Analyt. Chim. Acta 2003, 479, 59–75.CrossRefGoogle Scholar
  20. 20.
    Madl, J.; Rhode, S.; Stangl, H.; Stockinger, H.; Hinterdorfer, P.; Schutz, G. J.; Kada, G. Ultramicroscopy 2006, 106, 645–651.CrossRefGoogle Scholar
  21. 21.
    Kienberger, F.; Pastushenko, V.; Kada, G.; Gruber, H.; Riener, C.; Schindler, H.; Hinterdorfer, P. Single Molecules 2000, 1, 123–128.CrossRefADSGoogle Scholar
  22. 22.
    Kratky, O.; Porod, G. Recueil des Travaux Chimiques des Pays-Bas 1949, 68, 1106–1123.CrossRefGoogle Scholar
  23. 23.
    Rief, M.; Fernandez, J. M.; Gaub, H. E. Physical Review Letters 1998, 81, 4764.CrossRefADSGoogle Scholar
  24. 24.
    Marko, J. F.; Siggia, E. D. Macromolecules 1995, 28, 8759–8770.CrossRefADSGoogle Scholar
  25. 25.
    Odijk, T. Macromolecules 1995, 28, 7016–7018.CrossRefADSGoogle Scholar
  26. 26.
    Tskhovrebova, L.; Trinick, J.; Sleep, J.; Simmons, R. Nature 1997, 387, 308–312.CrossRefADSGoogle Scholar
  27. 27.
    Oesterhelt, F.; Rief, M.; Gaub, H. E. New J. Phys. 1999, 1, 1–11.CrossRefGoogle Scholar
  28. 28.
    Rixman, M. A.; Dean, D.; Ortiz, C. Langmuir 2003, 19, 9357–9372.CrossRefGoogle Scholar
  29. 29.
    Bell, G. I. Science 1978, 200, 618–27.CrossRefADSGoogle Scholar
  30. 30.
    Kramers, H. A. Physica 1940, 7, 284–304.zbMATHCrossRefADSMathSciNetGoogle Scholar
  31. 31.
    Jeffery, S.; Hoffmann, P. M.; Pethica, J. B.; Ramanujan, C.; Ozer, H. O.; Oral, A. Phys. Rev. B 2004, 70, 054114–8.CrossRefADSGoogle Scholar
  32. 32.
    Jeppesen, C.; Wong, J. Y.; Kuhl, T. L.; Israelachvili, J. N.; Mullah, N.; Zalipsky, S.; Marques, C. M. Science 2001, 293, 465–468.CrossRefGoogle Scholar
  33. 33.
    Friedsam, C.; Wehle, A. K.; Kuhner, F.; Gaub, H. E. J. Phys.-Cond. Matt. 2003, 15, S1709-S1723.CrossRefADSGoogle Scholar
  34. 34.
    Oesterhelt, F.; Rief, M.; Gaub, H. E. New J. Phys. 1999, 1, 1–11.CrossRefGoogle Scholar
  35. 35.
    Heymann, B.; Grubmuller, H. Chem. Phys. Lett. 1999, 307, 425–432.CrossRefADSGoogle Scholar
  36. 36.
    Zhang, Q. M.; Marszalek, P. E. Polymer 2006, 47, 2526–2532.CrossRefGoogle Scholar
  37. 37.
    Rief, M.; Gautel, M.; Oesterhelt, F.; Fernandez, J. M.; Gaub, H. E. Science 1997, 276, 1109–1112.CrossRefGoogle Scholar
  38. 38.
    Marszalek, P. E.; Lu, H.; Li, H. B.; Carrion-Vazquez, M.; Oberhauser, A. F.; Schulten, K.; Fernandez, J. M. Nature 1999, 402, 100–103.CrossRefADSGoogle Scholar
  39. 39.
    Carrion-Vazquez, M.; Li, H. B.; Lu, H.; Marszalek, P. E.; Oberhauser, A. F.; Fernandez, J. M. Nature Struct. Biol. 2003, 10, 738–743.CrossRefGoogle Scholar
  40. 40.
    Fernandez, J. M.; Li, H. Science 2004, 303, 1674–1678.CrossRefADSGoogle Scholar
  41. 41.
    Dietz, H.; Berkemeier, F.; Bertz, M.; Rief, M. Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 12724–12728.CrossRefADSGoogle Scholar
  42. 42.
    Muller, D. J.; Kessler, M.; Oesterhelt, F.; Moller, C.; Oesterhelt, D.; Gaub, H. Biophys. J. 2002, 83, 3578–3588.CrossRefGoogle Scholar
  43. 43.
    Dietz, H.; Rief, M. PNAS 2006, 103, 1244–1247.CrossRefADSGoogle Scholar
  44. 44.
    Holliger, P.; Prospero, T.; Winter, G. Proc. Natl. Acad. Sci. U.S.A. 1993, 6444–6448.Google Scholar
  45. 45.
    Baumgartner, W.; Hinterdorfer, P.; Ness, W.; Raab, A.; Vestweber, D.; Schindler, H.; Drenckhahn, D. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 4005–4010.CrossRefADSGoogle Scholar
  46. 46.
    Bonanni, B.; Kamruzzahan, A. S. M.; Bizzarri, A. R.; Rankl, C.; Gruber, H. J.; Hinterdorfer, P.; Cannistraro, S. Biophys. J. 2005, 89, 2783–2791.CrossRefGoogle Scholar
  47. 47.
    Sulchek, T. A.; Friddle, R. W.; Langry, K.; Lau, E. Y.; Albrecht, H.; Ratto, T. V.; DeNardo, S. J.; Colvin, M. E.; Noy, A. Proc. Natl. Acad. Sci. U.S.A. 2005, 102, 16638–16643.CrossRefADSGoogle Scholar
  48. 48.
    Leckband, D. E.; Israelachvili, J. N.; Schmitt, F. J.; Knoll, W. Science 1992, 255, 1419–1421.CrossRefADSGoogle Scholar
  49. 49.
    Ashkin, A. Proc. Natl. Acad. Sci. U.S.A. 1997, 94, 4853–4860.CrossRefADSGoogle Scholar
  50. 50.
    Ludwig, M.; Dettmann, W.; Gaub, H. E. Biophys. J. 1997, 72, 445–448.CrossRefGoogle Scholar
  51. 51.
    Grandbois, M.; Dettmann, W.; Benoit, M.; Gaub, H. E. J. Histochem. Cytochem. 2000, 48, 719–724.Google Scholar
  52. 52.
    Dupres, V.; Menozzi, F. D.; Locht, C.; Clare, B. H.; Abbott, N. L.; Cuenot, S.; Bompard, C.; Raze, D.; Dufrene, Y. F. Nature Methods 2005, 2, 515–520.CrossRefGoogle Scholar
  53. 53.
    Liu, F.; Arce, F. T.; Ramachandran, S.; Lal, R. J. Biol. Chem. 2006, 281, 23207–23217.CrossRefGoogle Scholar
  54. 54.
    Hinterdorfer, P.; Baumgartner, W.; Gruber, H. J.; Schilcher, K.; Schindler, H. PNAS 1996, 93, 3477–3481.CrossRefADSGoogle Scholar
  55. 55.
    Ebner, A.; Kienberger, F.; Kada, G.; Stroh, C. M.; Geretschlaeger, M.; Kamruzzahan, A. S. M.; Wildling, L.; Johnson, W. T.; Ashcroft, B.; Nelson, J. ChemPhysChem 2005, 6, 897–900.CrossRefGoogle Scholar
  56. 56.
    Stroh, C.; Wang, H.; Bash, R.; Ashcroft, B.; Nelson, J.; Gruber, H.; Lohr, D.; Lindsay, S. M.; Hinterdorfer, P. Proc. Natl. Acad. Sci. U.S.A. 2004, 101, 12503–12507.CrossRefADSGoogle Scholar
  57. 57.
    Hinterdorfer, P.; Dufrene, Y. F. Nature Methods 2006, 3, 347–355.CrossRefGoogle Scholar
  58. 58.
    Sulchek, T.; Hsieh, R.; Adams, J. D.; Yaralioglu, G. G.; Minne, S. C.; Quate, C. F.; Cleveland, J. P.; Atalar, A.; Adderton, D. M. Appl. Phys. Lett. 2000, 76, 1473–1475.CrossRefADSGoogle Scholar
  59. 59.
    Hobbs, J. K.; Vasilev, C.; Humphris, A. D. L. Analyst 2006, 131, 251–256.CrossRefADSGoogle Scholar
  60. 60.
    Mammen, M., Helmerson, K., Kishore, R., Choi, S. K., Phillips, W. D., and Whitesides G. M. Chem. & Biol. 1996, 3, 757–763.Google Scholar
  61. 61.
    Wells, J. A. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 1–6.CrossRefADSGoogle Scholar
  62. 62.
    Dower, S. K.; DeLisi, C.; Titus, J. A.; Segal, D. M. Biochemistry 1981, 20, 6326–6334.CrossRefGoogle Scholar
  63. 63.
    Kipriyanov, S. M.; Little, M.; Kropshofer, H.; Breitling, F.; Gotter, S.; Dubel, S. Protein Eng. 1996, 9, 203–211.CrossRefGoogle Scholar
  64. 64.
    Kortt, A. A.; Dolezal, O.; Power, B. E.; Hudson, P. J. Biomolec. Engin. 2001, 18, 95–108.CrossRefGoogle Scholar
  65. 65.
    Wong, S. S.; Joselevich, E.; Woolley, A. T.; Cheung, C. L.; Lieber, C. M. Nature 1998, 394, 52–55.CrossRefADSGoogle Scholar
  66. 66.
    Evans, E.; Ritchie, K. Biophys. J. 1997, 72, 1541–1555.CrossRefGoogle Scholar
  67. 67.
    Evans, E.; Williams, P. Dynamic Force Spectroscopy: II. Multiple Bonds, in Physics of Bio-Molecules and Cell; Springer and EDP Sciences: Heidelberg, 2002.Google Scholar
  68. 68.
    Han, T.; Williams, J. M.; Beebe, T. P. Analyt. Chim. Acta 1995, 307, 365–376.CrossRefGoogle Scholar
  69. 69.
    Williams, P. M. Analyt. Chim. Acta 2003, 479, 107–115.CrossRefGoogle Scholar
  70. 70.
    Jager, M.; Pluckthun, A. FEBS Letters 1999, 462, 307–312.CrossRefGoogle Scholar
  71. 71.
    Kienberger, F.; Kada, G.; Mueller, H.; Hinterdorfer, P. J. Molec. Biology 2005, 347, 597.CrossRefGoogle Scholar
  72. 72.
    Marcel, V.; Harry, B. Cell and Tissue Research 2000, V301, 133–142.Google Scholar
  73. 73.
    Evans, E.; Williams, P. In Physics of Bio-Molecules and Cells; Flyvbjerg, H., Jülicher, F., Ormos, P., David, F., Eds.; Springer and EDP Sciences: Heidelberg, 2002; Vol. 75, p 187–203.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Todd A. Sulchek
    • 1
  • Raymond W. Friddle
    • 2
  • Aleksandr Noy
    • 3
  1. 1.Chemistry, Materials and Life Sciences DirectorateLawrence Livermore National Laboratory, L-231LivermoreUSA
  2. 2.Chemistry, Materials and Life Sciences DirectorateLawrence Livermore National Laboratory, L-234LivermoreUSA
  3. 3.Chemistry, Materials and Life Sciences DirectorateLawrence Livermore National Laboratory, L-234LivermoreUSA

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